Formalin-fixed, paraffin-embedded skin tissues (3 samples of tuberculosis, 2 samples of tuberculids, and 10 samples of normal skins) were used for the immunohistochemical studies. For immunoperoxidase staining, sectioned tissues were treated for endogenous peroxidase inactivation (3% hydrogen peroxide). Then, the tissues were blocked. Specimens were then incubated overnight with primary antibody. Anti-HBD-2 antibody (Abcam, UK) was used at 1:100 dilution, anti-HBD-3 antibody (Novus Biological, USA) at 1:50 dilution, and anti-LL-37 antibody (Abcam, UK) was diluted to 1:500. Then, secondary antibody (anti-mouse/anti-rabbit antibody) was added sequentially for 30 min, followed by 3-amino-9-ethylcarbazole/hematoxylin color spectrum analysis. The result was observed by the use of DC 300F microscopic image analysis system (Leica Microsystems GmbH, Wetzlar, Germany), and the mean optical density values of three distinct groups were measured.
Anti hbd 3 antibody
Manufactured by Novus Biologicals
Sourced in United States
The Anti-HBD-3 antibody is a laboratory tool used for the detection and quantification of human beta-defensin 3 (HBD-3) in various samples. HBD-3 is a small antimicrobial peptide involved in innate immune responses. This antibody can be utilized in techniques such as ELISA, Western blotting, and immunohistochemistry to study the expression and localization of HBD-3 in biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Anti ll 37 antibody, by Abcam (2 mentions)
Anti hbd 2 antibody, by Abcam (2 mentions)
Polyvinylidene fluoride membrane, by Thermo Fisher Scientific (1 mentions)
Imagej software, by GE Healthcare (1 mentions)
Nupage 4 12 bis tris polyacrylamide gel, by Thermo Fisher Scientific (1 mentions)
Gentamycin solution, by Merck Group (1 mentions)
Mouse monoclonal anti α tubulin t5168, by Merck Group (1 mentions)
Rabbit monoclonal anti gapdh 14c10, by Cell Signaling Technology (1 mentions)
Shc002, by Merck Group (1 mentions)
3 protocols using anti hbd 3 antibody
1
Immunohistochemical Analysis of Skin Antimicrobial Peptides
2
Quantifying Skin Antimicrobial Peptides
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Skin tissues were homogenized in radioimmunoprecipitation assay buffer with phenylmethanesulfonyl fluoride on ice for 30 min, vibrated, and centrifuged. Total protein of the skin tissue lysates was evaluated by BCA protein assay (Fish Scientific, Fair Lawn, NJ, USA). Skin lysates containing 30 μg of crude skin tissue lysate protein were analyzed by NuPAGE 4–12% Bis-Tris polyacrylamide gels (Invitrogen, Carlsbad, CA, USA) and was transferred to polyvinylidene fluoride membranes (Invitrogen). The membranes were incubated overnight with the polyclonal rabbit anti-HBD-2 antibody (Abcam), anti-HBD-3 antibody (Novus Biological, USA), or anti-LL-37 antibody (Abcam) at 1:1000 dilution, respectively. The analysis was performed with chemiluminescence reagents. Protein expression was normalized to the quantity of beta-actin. The signal and grayscale values were visualized and analyzed by using ImageJ software (GE Healthcare Piscataway, NJ, USA), and grayscale value ratios were calculated.
3
Reagents and Plasmids for Cell Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
N-phosphonacetyl-l -aspartate (PALA, NSC224131) was obtained from the National Cancer Institute (NCI)/Division of Cancer Treatment and Diagnosis (DCTD)/Developmental Therapeutics Program (DTP) Open Chemical Repository (http:dtp.cancer.gov). Aquaphor® and Neosporin® were purchased from Target (Cleveland Heights, OH). Gentamycin solution was purchased from Sigma (St. Louis, MO). Rabbit monoclonal anti-CAD antibody (EP711Y) was purchased from Novus Biologicals (Littleton, CO). Rabbit polyclonal anti-RICK (H-300) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Rabbit monoclonal anti-GAPDH (14C10) was purchased from Cell Signaling Technologies (Danvers, MA). Mouse monoclonal anti-α-tubulin (T5168) was purchased from Sigma. Antibodies against LL-37 (sc-166770) and HBD2 (sc-20798) were purchased from Santa Cruz Biotechnology. Anti-HBD3 antibody was purchased from Novus Biologicals (NB200-117). The plasmids, pcDNA3-HA-NOD1, pcDNA3-HA-NOD2, pBVIII-Luc, and pCMV-βgal were gifts of Gabriel Nuñez (University of Michigan) and have been previously described56 (link). MISSION short hairpin RNA (shRNA) constructs targeting CAD (NM_004341.3-6910s21c1), NOD2 (NM_022162.1-2959s1c1), RIP2 (NM_003821.5-2364s21c1), and non-targeting shRNA control (SHC002) were purchased from Sigma.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!